This invention relates to a novel estrogen receptor and to the polynucleotide sequences encoding this receptor. This invention also relates to methods for identifying ligands which bind to this receptor, to the ligands so identified, and to pharmaceutical compositions comprising such ligands. This invention also relates to pharmaceutical compositions useful for treating or preventing estrogen receptor mediated diseases or conditions.
Nuclear receptors are a large class of proteins that are responsible for the regulation of complex cellular events including cell differentiation, homeostasis, the growth and functioning of various organs and tissues, and transcription. It is believed that nuclear receptors function by transducing extracellular chemical signals from hormones into a transcriptional response.
Estrogen receptors are a subclass of the larger nuclear receptor class. The estrogen receptors are proteins that are responsive to estrogen and estrogen-like molecules. Estrogen receptors are believed to play an important role in the mammalian endocrine system, the reproductive organs, breast tissue, bone tissue, and the vascular system, and are believed to be involved in the development and progression of various disease states such as abnormal bone resorption, cardiovascular disease, cancer, and central nervous system disorders. It is believed that various disease states and conditions can be treated or prevented by the development of appropriate ligands, i.e. drugs, for modifying the activity of estrogen receptors. Consequently there is a need to identify estrogen receptors and their mode of action and to also identify ligands for modifying the action of these receptors.
At least two distinct types of estrogen receptors have been reported. An estrogen receptor having 595 amino acids is disclosed in Green, S. et al., Nature, 320, pp. 134-139 (1986) and Greene, G. L. et al., Science, 231, pp. 1150-1154 (1986), both of which are incorporated by reference herein in their entirety. These references also disclose the corresponding DNA sequences for the receptor.
The other reported type of estrogen receptor has been disclosed by two research groups and has been designated xe2x80x9cxcex2xe2x80x9d (beta). One research group discloses a 485 amino acid xcex2 preceptor that is obtained from rat, human, and mouse sources, as well as the corresponding DNA sequences. See PCT application No. WO 97/09348, to Kuiper, G. G. J. M. et al., published Mar. 13, 1997, which is incorporated by reference herein in its entirety. The second research group discloses a similar estrogen receptor containing 483 amino acids. The corresponding DNA sequence is also disclosed. See Mosselman, S. et al., ERxcex2: identification and characterization of a novel human estrogen receptor, FEBS Letters, 392, pp. 49-53 (1996), which is incorporated by reference herein in its entirety.
In the present invention, a novel estrogen receptor having 548 amino acid units, and that is distinct from the disclosed 595 amino acid, 485 amino acid, and 483 amino acid estrogen receptors, has been identified and isolated from human tissue. It is believed that this novel estrogen receptor plays a key role in mammalian physiology. This novel estrogen receptor is an important research tool for identifying and designing ligands for use in pharmaceutical compositions for treating and/or preventing a wide range of estrogen receptor mediated diseases or conditions.
It is therefore an object of the present invention to provide a novel isolated estrogen receptor.
It is another object of the present invention to provide the amino acid sequence of a novel estrogen receptor.
It is another object of the present invention to provide the polynucleotide sequence encoding a novel estrogen receptor.
It is another object of the present invention to provide methods for isolating a novel estrogen receptor.
It is another object of the present invention to provide ligands capable of binding to a novel estrogen receptor.
It is another object of the present invention to provide pharmaceutical compositions comprising ligands capable of binding to a novel estrogen receptor.
It is another object of the present invention to provide methods for treating and/or preventing estrogen receptor mediated diseases or conditions.
These and other objects will become readily apparent from the detailed description which follows.
The present invention relates to an isolated estrogen receptor comprising the amino acid sequence of FIG. 1 (which also corresponds to SEQ ID NO: 1).
In further embodiments, the present invention relates to an isolated estrogen receptor having an amino acid sequence that is substantially similar to the amino acid sequence of FIG. 1 (SEQ ID NO: 1), wherein the estrogen receptor comprises at least 531 amino acids.
In further embodiments, the present invention relates to an isolated estrogen receptor comprising at least 531 amino acids and having substantially the same ligand binding properties or substantially the same DNA binding properties as the estrogen receptor of FIG. 1 (SEQ ID NO: 1).
In further embodiments, the present invention relates to an isolated estrogen receptor that is derived from mammalian cells, preferably human cells.
In further embodiments, the present invention relates to an isolated polynucleotide encoding the estrogen receptor having the amino acid sequence of FIG. 1 (SEQ ID NO: 1).
In further embodiments, the present invention relates to an isolated polynucleotide which is a DNA, a cDNA, or an RNA.
In further embodiments, the present invention relates to an isolated polynucleotide which hydridizes to and is complementary to the polynucleotide encoding the estrogen receptor having the amino acid sequence of FIG. 1 (SEQ ID NO: 1).
In further embodiments, the present invention relates to an isolated polynucleotide comprising a polynucleotide encoding a mature polypeptide encoded by the estrogen receptor polynucleotide contained in an ATCC Deposit selected from the group consisting of ATCC Deposit No. 209238, ATCC Deposit No. 209239, and ATCC Deposit No. 209240.
In further embodiments, the present invention relates to an isolated polynucleotide comprising the nucleotide sequence of FIG. 2 (which also corresponds to SEQ ID NO: 2).
In further embodiments, the present invention relates to an isolated polynucleotide which hybridizes to and is complementary to the polynucleotide of FIG. 2 (SEQ ID NO: 2), wherein said polynucleotide comprises at least 1593 nucleotides.
In further embodiments, the present invention relates to a vector containing the DNA.
In further embodiments, the present invention relates to a host cell transformed or transfected with the vector of the present invention.
In further embodiments, the present invention relates to a method for producing an estrogen receptor of the present invention.
In further embodiments, the present invention relates to a method for determining whether a ligand can bind to the estrogen receptor of the present invention.
In further embodiments, the present invention relates to a ligand detected by the methods of the present invention.
In further embodiments, the present invention relates to a pharmaceutical composition comprising a ligand of the present invention.
In further embodiments, the present invention relates to a method for treating or preventing an estrogen receptor mediated disease or condition by administering an effective amount of a pharmaceutical composition of the present invention.
The deposits referred to herein will be maintained under the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the purposes of Patent Procedure. These deposits are provided merely as a convenience and are not an admission that a deposit is required under 35 USC xc2xa7112. The sequence of the polynucleotides contained in the deposited materials, as well as the amino acid sequence of the polypeptides encoded thereby, are incorporated herein by reference in their entirety and are controlling in the event of any conflict with the description of the sequences herein. A license may be required to make, use or sell the deposited materials, and no such license is hereby granted.
All percentages and ratios used herein, unless otherwise indicated, are by weight. The invention hereof can comprise, consist of, or consist essentially of the essential as well as optional ingredients, components, and methods described herein.